Isothermal DNA amplification facilitates the identification of a broad spectrum of bacteria,fungi and protozoa in <Emphasis Type="Italic">Eleutherococcus</Emphasis> sp. plant tissue cultures |
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Authors: | Peter Müller Matthias Döring |
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Institution: | 1. UMR DIAPC, IRD/CIRAD Palm Development Group, IRD, 34394, Montpellier, France 5. Faculté des Sciences et Techniques, Laboratoire de Biotechnologies Végétales, Université Cheikh Anta Diop, BP 5005, Dakar, Senegal 7. Genetic Resource Center, International Institutes of Tropical Agriculture, PMB 5320, Ibadan, Nigeria 2. UMR DIAPC, IRD/CIRAD Palm Development Group, CIRAD, 34394, Montpellier, France 3. UMR BGPI, Microscopie Electronique, CIRAD, 34398, Montpellier, France 4. Macalester College, St. Paul, MN, 55105, USA 6. UMR DAP, PHIV, CIRAD, 34398, Montpellier, France
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Abstract: | In vitro cultures of Eleutherococcus sieboldianus originating from surface sterilized leaf explants were found to be associated with several microorganisms. The associations
included bacteria, fungi and protozoa within the rhizosphere and inside root hairs. To determine if this phenomenon is unique
to this species, plant tissue cultures of E. gracilistylus and E. senticosus were included in our studies for comparison. A methodology consisting of isothermal amplification, cloning and sequencing
was established for analysing 16S ribosomal DNA of cultivated and non-cultivated bacteria from different tissue types. The same methodology was used to obtain
internal transcribed spacer regions and 18S regions of fungal and protozoan rDNA. Comparative analyses of sequencing data resulted in the identification of various genera
within the Firmicutes and γ-proteobacteria kingdoms and a broad spectrum of fungal genera related to several uncultured fungi.
In addition, amoebal and chrysophyte species were detected. Most of the species were identified in different plant organs
and in in vitro culture cell types indicating the microorganisms are systemically distributed. The presence of identical microorganisms
in different plant species argues for an evolutionary long-lasting and stable association between the plant genus and the
microinhabitants. |
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Keywords: | Acanthopanax (Araliaceae) Root hair endophytes Non-cultured microorganisms Internal transcribed spacer region (ITS) Phi-29 DNA polymerase and isothermal DNA amplification |
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